CXCR4-siRNA重组腺病毒载体沉默人卵巢癌细胞株CXCR4基因表达的研究  

Study on the expression of CXCR4 gene silenced by CXCR4-siRNA recombinant adenoviral vector in human ovarian cancer cells

作　　者：刘瑜[1] 袁瑞[1] 

机构地区：[1]重庆医科大学附属第一医院妇产科,重庆400016

出　　处：《重庆医科大学学报》2011年第10期1168-1171,共4页Journal of Chongqing Medical University

基　　金：重庆市自然科学基金计划资助项目(编号:CSTC;2008BB5239)

摘　　要：目的:研究CXCR4-siRNA重组腺病毒载体对人卵巢癌细胞株SKOV3的CXCR4基因表达的影响。方法:PacⅠ酶切CXCR4-siRNA重组腺病毒载体使其线性化,应用HEK293细胞大量包装扩增病毒,收集病毒液,转染人卵巢癌细胞株SKOV3。细胞分为3组:siRNA组,转染空病毒载体的阴性对照组,细胞不做任何处理的空白对照组。通过PCR、Western blot、免疫组化检测CXCR4基因沉默效果。结果:包装扩增后病毒滴度为6×108 PFU/ml,荧光显微镜下可观察到SKOV3细胞中绿色荧光蛋白的大量表达,重组腺病毒能够有效抑制SKOV3细胞中CXCR4的表达。结论:CXCR4-siRNA重组腺病毒载体可高效转染靶细胞和沉默CXCR4基因的表达,为RNA干扰技术应用于卵巢癌的临床基因治疗奠定基础。Objective：To study the effect of the CXCR4-siRNA recombinant adenoviral vector on the expression of CXCR4 gene in SKOV3 cells.Methods：Recombinant adenoviral vector was linearized with PacⅠ.Then it was repeatedly packaged and amplified in HEK293 cells and transfected into SKOV3 cells.The SKOV3 cells were divided into three groups：siRNA group with adenovirus carrying RNA interference,negative group with vacant adenovirus vector,and control group without any infection.The expression level of CXCR4 gene in SKOV3 cells was measured by RT-PCR,Western blot and immunocytochemistry analysis.Results：The viral titers were 6×108 PFU/ml.The green fluorescent protein expressed highly in SKOV3 cells under fluorescent microscope.The CXCR4 gene was greatly inhibited after infection with CXCR4-siRNA recombinant adenoviral vector.Conclusion：The CXCR4-siRNA recombinant adenoviral vector can efficiently transfect targeted cells and silence the expression of CXCR4 gene,which would lay a foundation for the further study on clinical gene therapy of RNAi technology in ovarian cancer.

关 键 词：CXCR4 RNA干扰 重组腺病毒载体 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]